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Nightmare Moon is set to return every century (rather than every 1000 years; it's an adaptaion, after all). It's during that time the ponies that would represent the six Elements of Harmony: Honesty, Kindness, Laughter, Generosity, Loyalty and Magic would unite together in an attempt to defeat Nightmare Moon and prevent eternal night from happening. After her last defeat, Equestria enjoyed four years of peace, until Trixie, using dark magic, had resurrected the spirit of Nightmare Moon and began corrupting ponies, who wielded the Elements of Harmony, and purging their memories. Rarity, Rainbow Dash, Pinkie Pie and Fluttershy (altough only partially) were turned into Nightmare Moon's servants. Applejack and Twilight Sparkle avoided the corruption, only because Applejack was searching for Rainbow Dash, the pony who defeated Nightmare Moon four years ago, and vanished.However, she did not escaped the memory purge. Twilight wasn't corrupted at all, as she decided to enter a deep slumber after Nightmare's Moon's defeat at her hooves, to contain her magical potential and keep it from going out of control.

It all goes to a waste when Trixie's plan is enacted. With Princess Celestia and Luna banished to the sun and the moon respectively, it's up to Twilight Sparkle to save Equestria from the eternal night. Assisted by her familiar, Spike, she goes on her crusade against Nightmare Moon.